This is a competing renewal requesting funds for the continuance of the short course "Methods in Human ES Cell Research" formerly known as "Current Protocols in Stem Cell Biology" which has been offered annually at The Jackson Laboratory since 2002. This course will continue to be held in The Jackson Laboratory's Applied Genomics Training Center in early August of each year. This course will continue to bring together some of the leading experts on ES cell technology to train students in the successful culture, maintenance and manipulation of ES cells. Specifically, this course will cover: 1) The preparation of media, including necessary quality control; 2) The preparation of feeder layers; 3) ES cell passaging techniques; 4) ES cell cryopreservation; 5) ES cell transfection techniques; 6) FACs analysis of ES cells and their differentiated derivatives; 7) Embryoid body formation; 8) Neural Differentiation of ES cells; 9) Hematopoietic Differentiation of ES cells; and 10) Cardiac Differentiation of ES cells. The Course will utilize only human ES cell lines registered with the NIH, specifically UC06, WA09 and ES03. One major long term goal for this course will be to share and improve standard protocols, and eventually to publish a techniques manual, similar in style to the popular "Manipulating the Mouse Embryo" manual edited by Hogan et al. In fact, the course syllabus currently contains a compendium of protocols, reagent lists and research papers, providing stem cell researchers with a valuable reference resource. These materials are available upon request from The Jackson Laboratory. These aims will be accomplished by offering an intensive 5-day course to 16 participants chosen for their outstanding research potential. In 2006, the course will be held on August 6-11 at The Jackson Laboratory in Bar Harbor, Maine, and will be immediately preceded by the 2nd Annual International Stem Cell Initiative Workshop at which human ES cell researchers from the major ES cell research centers around the world will meet to discuss the outcome of experiments carried out using shared reagents and methods. The course will be immediately followed by a companion symposium "Stem Cells on Land and at Sea", also held annually, at The Mount Desert Island Biological Laboratory which examines broader issues in comparative stem cell biology. Students in the course are encouraged to attend these companion events free of charge. Relevance to Public Health: The current major bottleneck to human ES cells becoming a ubiquitous research tool in the biomedical research community continues to be the scarcity of the hands-on training necessary to learn how to culture these sensitive cells. The course proposed in this application will continue to provide hands-on training for investigators in how to culture, manipulate, and differentiate human ES cells in vitro. [unreadable] [unreadable] [unreadable]